Each year in the United States at least 1.4 million people seek medical attention for a traumatic brain injury (TBI). Between 75% and 90% of these are mild or moderate injuries. These can inflict profound and persistent impairment of physical, cognitive, and psychosocial functioning. The physical, social and economic sequelae of TBI can persist for the victim's entire life, a burden that is borne by the individual, their family, and by society. However, mild and moderate TBI is challenging to diagnose. Conventional methods such as imaging and neurological examination are often insensitive to mild injury. There is no blood- based diagnostic tool to aid clinicians in the diagnosis of mild and moderate TBI. However, such a rapid, reliable, and sensitive tool would greatly improve diagnosis and triage of TBI patients, would more accurately guide treatment and management options, and therefore would improve patient outcome while reducing management costs. Banyan Biomarkers Inc. has been recognized as a pioneer in the development of ultra-sensitive brain injury biomarker diagnostic assays. We have the commitment to providing such a tool to the marketplace, based on proprietary TBI biomarkers. Previously, in our Phase I SBIR project, a sensitive ELISA for quantifying one brain injury biomarker (BA0123) in CSF was successfully developed, and proof-of-concept data confirming that BA0123 is increased in CSF following brain injury was obtained from preclinical studies. Subsequently, a second complementary neuro-biomarker (BA0252) was discovered and a specific ELISA assay for its sensitive detection was developed. Proof-of-concept data to support the hypothesis that both biomarkers are increased in CSF and serum of human patients with severe TBI was generated and preliminary data show detection of these markers also in mild and moderate TBI patients. The data supporting the development of both biomarkers as tools to aid in the diagnosis of TBI are very compelling. In Phase II of this project a commercializable ELISA assay kit for both biomarkers (BA0123 and BA0252) will be developed. Assay components (antibodies and antigens) will be produced in abundant supply and under quality assured conditions. Feasibility data will be collected from clinical studies of severe, moderate, and mild TBI that will allow correlation of biomarker levels to clinical measures of injury severity, disease progression, and outcome. The accuracy with which the biomarkers can discriminate magnitude of injury, and patients that are likely to have poor progression and outcome will be evaluated. This analysis will determine the sensitivity and specificity of markers BA0123 and BA0252 when used singularly or in combination. These data will be used to support a pre-IDE application to the FDA. The deliverables of this Phase II project are two quality-assured ELISA assays for the brain injury biomarkers establishing the foundation toward their commercialization. These assays will be used to analyze clinical samples, further confirming the use of the biomarkers as tools to aid in the diagnosis of severe, mild and moderate TBI. [unreadable] Each year in the United States at least 1.4 million people sustain a TBI, of which about 50,000 die, 235,000 are hospitalized, and 1.1 million are treated and released from an emergency department. Current methods used to assess the existence and severity of brain injury, based on imaging techniques such as CT scanning or MRI, are expensive, not consistently reliable, not universally available, and are slow to produce results. Banyan Biomarkers is developing rapid, reliable, sensitive, and economical blood tests for brain injury that would greatly improve diagnostic and triage capabilities, would more accurately guide treatment options, and therefore would improve patient outcome. [unreadable] [unreadable] [unreadable]